| Objective:Gastric cancer is one of the most common malignant tumors in the world,its pathogenesis is unknown.In China,most patients are in advanced stage with poor prognosis.Various long-chain non-coding RNA are involved in the development,invasion and metastasis of gastric cancer.Long non-protein coding RNA(lncRNA)is a non-protein-encoding RNA that is defined as a non-coding RNA greater than 200 nt in length.Studies have shown that lncRNA can participate in the biological processes of gastric cancer,including chromatin remodeling,cell differentiation and immune response.LINC01133 is a newly discovered lncRNA,and its role in the development of gastric cancer remains unclear.This project mainly studied the expression of LINC01133 in gastric cancer tissues and the effects on the proliferation,cycle,apoptosis,metastasis and invasion of gastric cancer cells cultured in vitro.Method:1.The expression of LINC01133 in 30 cases of gastric cancer tissues and their adjacent tissues were detected by qRT-PCR.The correlation between the differential expression of LINC01133 and clinicopathological features were analyzed.2.Using qRT-PCR to detect the expression level of LINC01133 in normal gastric mucosal epithelial cells GES-1 and human gastric cancer cell lines MGC-803 and BGC-823.3,LINC01133 overexpression vector was constructed and transfected into MGC-803 human gastric cancer cell line,LINC01133 interference vector was constructed and transfected into BGC-823 human gastric cancer cell line.The cell experiments were further grouped into: 1)NC group(MGC-803 transfected overexpression vector).2)LINC01133 group(MGC-803 transfected with LINCO1133-pcDNA3.1 plasmid group).3)sh-NC group(BGC-823 transfectioninterference empty vector group).4)sh-LINC01133-1 group(BGC-823 transfected siRNA-LINC01133 group).Cell proliferation was detected by CCK-8 method,cell cycle and apoptosis were detected by flow cytometry,cell migration was detected by wound staining assay,and cell invasion was detected by transwell assay.Results:1.Among the 30 gastric cancer tissues and their matched paracancerous tissues,the expression level of LINC01133 in gastric cancer tissues was significantly lower than that in adjacent normal tissues,P<0.001.2.There was no significant difference in the expression level of LINC01133 between GES-1 and BGC-823,P>0.05.The expression of LINC01133 in MGC-803 was lower than that in GES-1 and BGC-823 cells,P <0.001.3.Using CCK-8 method to detect MGC-803 with high expression of LINC01133,the cell survival rate of LINC01133 group(81.11%±0.82%)was significantly lower than that of NC group(95.68%±0.92%),the difference was significant,P <0.05.Using CCK-8 method to detect BGC-823,which interfered with the expression of LINC01133.The cell viability of sh-LINC01133-1 group(62.44%± 3.62%)was significantly higher than that of sh-NC group(60.43%±0.31%),P <0.05.4.MGC-803 with high expression of LINC01133 was detected by flow cytometry.The cells in LINC01133 group were up-regulated in G2 phase compared with NC group.Flow cytometry detected BGC-823 which interfered with LINC01133.The proportion of cells in sh-LINC01133-1 group was significantly lower than that in sh-NC group.5,MGC-803 with high expression of LINC01133 was detected by flow cytometry.The apoptosis rate of the NC group(24.25%±3.69%)was significantly lower than that of the LINC01133 group(90.06%±5.16%),P<0.05.Flow cytometry detected BGC-823,which interfered with LINC01133.The apoptosis rate of sh-NC group(65.03%±0.99%)was higher than that of sh-LINC01133-1(60.23%±2.51%).<0.05.6.Wound test was used to detect MGC-803 with high expression of LINC01133.The migration rate of cells in LINC01133 group(1.61 ± 0.16,?m/h)was lower thanthat in NC group(2.51 ± 0.11,?m/h),P<0.05.Wound test was used to detect BGC-823 interference with LINC01133.The migration rate of sh-LINC0113-1 group(3.09 ± 0.14,?m/h)was higher than that of sh-NC group(1.64 ± 0.44,?m/h),P<0.05.7.Transwell invasion assay was used to detect MGC-803 with high expression of LINC01133.The invasion of LINC01133 cells(0.1696 ± 0.0006)was lower than that of NC group(0.2083 ± 0.0003),P<0.05.After transwell invasion assay to detect BGC-823 in the LINC01133 group,the invasive quantification of the sh-LINC01133-1 group(0.2066 ± 0.0006)was higher than that of the sh-NC group(0.1831 ± 0.0012),P < 0.05.Conclusion:Low expression of LINC01133 in gastric cancer tissues,which can promote the proliferation,migration,invasion and inhibition of apoptosis of gastric cancer cells.LINC01133 plays a tumor suppressor gene role in the development of gastric cancer and is expected to become a new target in the study of gastric cancer. |
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